Effect of irisin on the repair capacity of tendon stem/progenitor cells in diabetic rats
Objective To explore the effects of irisin on the repair ability of tendon stem/progenitor cells in diabetes mellitus tendinopathy rats.Methods TSPCs were divided into control(complete medium),high glucose(complete medium+glucose 4.5 g/L)and irisin groups(complete medium+glucose 4.5 g/L+500 ng/ml Lrisin),the effects of irisin on the proliferation and migration of TSPCs were evaluated by cell CCK-8 assay,scratch assay,and Transwell assay.Meanwhile,Western blot method and immunofluorescence staining were used to investigate the effect of irisin on the expression of TSPCs differentiation-related proteins and its potential molecular mechanism.Finally,24 8-week-old male SD rats with body weight(300±20)g were divided into three groups by random number table method.The control(healthy rats+intraperitoneal saline),diabetes(diabetic rats+intraperitonealsaline)and irisin groups(diabetic rats+intraperitoneal irisin)were set up,the diabetic rat model was established,and irisinwas given to intervene,and the achilles tendon tissues of each group were stained to evaluate the effect of irisin on the differentiation ability of TSPC in vivo.Results The results of CCK8 experiment showed that TSPC proliferative activity decreased in the high glucose group compared with the control group(P＜0.01).The proliferative activity of TSPC in irisin group was higher than that in high glucose group(P＜0.01).Cell scratch and Transwell tests showed that TSPC migration ability was lower in the hyperglycemic group than in the control group(P＜0.01).The TSPC migration ability of irisin group was stronger than that of high glucose group(P＜0.01).Western blot and immunofluorescence staining showed that the expressions of Tnmd,Collagen Ⅰ,and SCX in the hyperglycemic group were lower than those in the control group,while the expressions of Sox9 and MMP-13 were increased(P＜0.01).Compared with high glucose group,the expressions of Tnmd,Collagen Ⅰ,and SCX were increased,while the expressions of Sox9 and MMP-13 were decreased in irisin group(P＜0.05).Compared with the control group,the expression of P-PI3K and P-Akt in high glucose group decreased(P＜0.01).The expressions of P-PI3K and P-Akt in irisin group were higher than those in hyperglycemic group(P＜0.05).The in vivo experimental results showed that compared with diabetes group,the degree of Achilles tendon lesions in irisin group was significantly reduced,Tnmd expression was increased,and Sox9 expression was decreased(P＜0.01).Conclusion Irisin can enhance the proliferation and migration of TSPCs in high-glucose microenvironment,and inhibit its misdifferentiation,thus delaying the progression of achilles tendinopathy in diabetic rats.
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